LOCUS       sequence-258381-        7245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTKMV2-pTDH3_FdeR-N_2NLS_tPGK1-pGPM1-fdeO_mcherry_tTDH1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7245)
  AUTHORS   Wang R, Cress BF, Yang Z, Hordines JC 3rd, Zhao S, Jung GY, Wang Z, 
            Koffas MAG
  TITLE     Design and Characterization of Biosensors for the Screening of 
            Modular Assembled Naringenin Biosynthetic Library in Saccharomyces 
            cerevisiae.
  JOURNAL   ACS Synth Biol. 2019 Sep 20;8(9):2121-2130. doi: 
            10.1021/acssynbio.9b00212. Epub 2019 Aug 28.
  PUBMED    31433622
REFERENCE   2  (bases 1 to 7245)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.9b00212"; journalName: "ACS Synth Biol"; date: 
            "2019-09-20- 20"; volume: "8"; issue: "9"; pages: "2121-2130"
COMMENT     Sequence Label: S3_004_026_MV2
FEATURES             Location/Qualifiers
     source          1..7245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(40..855)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(40..855)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     763..782
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     847..866
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1080..1668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1569..1588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1837..2503
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     2373..2396
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             3446..3466
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3470..3490
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5359..6162
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      6245..7227
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 ccgagcggcc gcgtgttaca accaattaac caattctgat tagaaaaact catcgagcat
       61 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
      121 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
      181 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
      241 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
      301 aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
      361 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga ggcgaaatac
      421 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
      481 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
      541 tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
      601 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
      661 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
      721 cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
      781 cccatataaa tcagcatcca tgttggaatt taatcgcggc ctggagcaag acgtttcccg
      841 ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
      901 tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg
      961 gctttgttga ataaatcgaa cttttgctga gttgaaggat cagtcatgac caaaatccct
     1021 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1081 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1141 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1201 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     1261 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     1321 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     1381 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     1441 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     1501 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     1561 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     1621 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     1681 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     1741 ttatcccctg attctgtgga taaccgtgcg gccgcccctg aattcgcatc tagactgaac
     1801 tggccgataa ttgcagacga acgcagttcg agtttatcat tatcaatact gccatttcaa
     1861 agaatacgta aataattaat agtagtgatt ttcctaactt tatttagtca aaaaattagc
     1921 cttttaattc tgctgtaacc cgtacatgcc caaaataggg ggcgggttac acagaatata
     1981 taacatcgta ggtgtctggg tgaacagttt attcctggca tccactaaat ataatggagc
     2041 ccgcttttta agctggcatc cagaaaaaaa aagaatccca gcaccaaaat attgttttct
     2101 tcaccaacca tcagttcata ggtccattct cttagcgcaa ctacagagaa caggggcaca
     2161 aacaggcaaa aaacgggcac aacctcaatg gagtgatgca acctgcctgg agtaaatgat
     2221 gacacaaggc aattgaccca cgcatgtatc tatctcattt tcttacacct tctattacct
     2281 tctgctctct ctgatttgga aaaagctgaa aaaaaaggtt gaaaccagtt ccctgaaatt
     2341 attcccctac ttgactaata agtatataaa gacggtaggt attgattgta attctgtaaa
     2401 tctatttctt aaacttctta aattctactt ttatagttag tctttttttt agttttaaaa
     2461 caccaagaac ttagtttcga ataaacacac ataaacaaac aaaagatcta tgcgtttcaa
     2521 caagctcgac ctcaatcttc tggtcgccct ggatgcactg ctcacggaga tgagcatcag
     2581 ccgcgccgcc gaaaagatcc atctgagcca gtcggccatg agcaatgccc tggcgcggct
     2641 gcgcgagtat ttcgatgatg aattgctgat ccaggtgggc cggcgcatgg agcccacgcc
     2701 gcgcgccgag gtgctcaagg atgcggtgca tgatgtgctg cggcgtatcg atggctccat
     2761 cgcggcgctg ccggccttcg tgccggccga gtccacgcgc gagtttcgca tctcggtttc
     2821 ggactttacg ctctccgtcc tcatcccccg ggtgctggcg cgcgcgcacg ccgagggcaa
     2881 gcacatccgc tttgccctga tgccgcaggt gcaagacccg acccgctcgc tggatcgggc
     2941 cgaggtggac ctgctggtct tgccgcagga attctgcacg cccgatcatc ctgccgaaga
     3001 ggtcttccgc gaacggcatg tctgcgtggt ctggcgcgac agtgcgctgg cgcaaggcga
     3061 gctgacgctg gaacgctaca tggcctcagg ccatgtggtg atggtgccgc ctggggccaa
     3121 tgcgtcgtcg gtggaggcgt ggatggccag gaagctgggc tttgcgcgcc gggtggaagt
     3181 gaccagcttc agcttcgctt ctgcgctggc gctggtacag gggacggacc gcatcgccac
     3241 ggtgcatgcc cggctggcgc agctgctggc tccgcaatgg ccggtggtga tcaaggagag
     3301 tccgctgtcg ctgggcgaga tgcggcagat gatgcagtgg catcgctacc gcagcaatga
     3361 tcctggcatc cagtggctgc gtcgggtgtt tctggagagt gcgcaggaga tggatgcggc
     3421 gctgccaggc atctgcggat ccgatccaaa aaagaagaga aaggtagatc caaaaaagaa
     3481 gagaaaggta taactcgagt ggcattgaat tgaattgaaa tcgatagatc aatttttttc
     3541 ttttctcttt ccccatcctt tacgctaaaa taatagttta ttttattttt tgaatatttt
     3601 ttatttatat acgtatatat agactattat ttatctttta atgattatta agatttttat
     3661 taaaaaaaaa ttcgctcctc ttttaatgcc tttatgcagt ttttttttcc cattcgatat
     3721 ttctatgtgc tgccaaacca gatgtcaaca cagctacaac gaagcttcgt cttttcttga
     3781 agaaaagagg aagctcgcta aatgggattc cactttccgt tccctgccag ctgatggaaa
     3841 aaggttagtg gaacgatgaa gaataaaaag agagatccac tgaggtgaaa tttcagctga
     3901 cagcgagttt catgatcgtg atgaacaatg gtaacgagtt gtggctgttg ccagggaggg
     3961 tggttctcac caaatcgcta cttgggtttg agattggtaa aattgattgt ttggatggca
     4021 atcatctatg gaatggatat tatataacaa agaagaaata atgaactgat tctcttcctc
     4081 cttcttgtcc tttcttaatt ctgttgtaat taccttcctt tgtaattttt tttgtaatta
     4141 ttcttcttaa taatccaaac aaacacacat attacaataa gatctatggt ttcaaagggc
     4201 gaagaagaca atatggctat tattaaggaa ttcatgagat tcaaggtcca catggaaggt
     4261 tctgtcaacg gtcacgaatt cgaaattgaa ggtgaaggtg aaggtagacc atacgaaggt
     4321 acccaaaccg ctaagttgaa ggtcaccaag ggtggtccat tgccattcgc ttgggatatt
     4381 ttgtctccac aattcatgta cggttctaag gcttacgtca agcacccagc tgatattcca
     4441 gattacttga agttgtcttt cccagaaggt ttcaagtggg aaagagtcat gaacttcgaa
     4501 gatggtggtg tcgtcaccgt cacccaagat tcttctttgc aagatggtga attcatttac
     4561 aaggtcaagt tgagaggtac caacttccca tctgatggtc cagtcatgca aaagaagacc
     4621 atgggttggg aagcttcttc tgaaagaatg tacccagaag atggtgcttt gaagggtgaa
     4681 attaagcaaa gattgaagtt gaaggatggt ggtcactacg atgctgaagt caagaccacc
     4741 tacaaggcta agaagccagt ccaattgcca ggtgcttaca acgtcaacat taagttggat
     4801 attacctctc acaacgaaga ttacaccatt gtcgaacaat acgaaagagc tgaaggtaga
     4861 cactctaccg gtggtatgga tgaattgtac aagtaaatcc taactcgaga taaagcaatc
     4921 ttgatgagga taatgatttt tttttgaata tacataaata ctaccgtttt tctgctagat
     4981 tttgtgatga cgtaaataag tacatattac tttttaagcc aagacaagat taagcattaa
     5041 ctttaccctt ttctttctaa gtttcaatat tagttatcac tgtttaaaag ttatggcgag
     5101 aacgtcggcg gttaaaatat attaccctga acggctgagc agttacagag atgttacgaa
     5161 ccactagtgc actgcagtac acggtttcct tgaaattttt ttgattcggt aatctccgaa
     5221 cagaaggaag aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt
     5281 gttgaagaaa catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacgtgc
     5341 aggaaacgaa gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct
     5401 agtcctgttg ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct
     5461 tcattggatg ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa
     5521 atttgtttac taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt
     5581 aagccgctaa aggcattatc cgccaagtac aattttttac tcttcgagga cagaaaattt
     5641 gctgacattg gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa
     5701 tgggcagaca ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag
     5761 caggcggcag aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca
     5821 tgcaagggct ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc
     5881 gacaaagatt ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt
     5941 tacgattggt tgattatgac acccggtgtg ggtttagatg acaagggaga tgcattgggt
     6001 caacagtata gaaccgtgga tgatgtggtt tctacaggat ctgacattat tattgttgga
     6061 agaggactat ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca
     6121 ggctgggaag catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta
     6181 aatgcatgta tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc
     6241 gagtaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat
     6301 ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct
     6361 attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg
     6421 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga
     6481 gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga
     6541 gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat
     6601 aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact
     6661 ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt
     6721 actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac
     6781 cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg
     6841 tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt
     6901 tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc
     6961 taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc
     7021 aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa
     7081 gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac
     7141 agtccggtgc gtttttggtt ttttgaaagt gcgtcatcag agcgcttttg gttttcaaaa
     7201 gcgctctgaa gttcctatac tttctagcta gagaatagga acttc
//
